Effects of calmodulin antagonists on sodium-dependent high-affinity choline uptake

The effects of calmodulin (CaM) antagonists were investigated on the sodium-dependent high-affinity choline uptake (SDHACU) as assessed by the specific binding of [3H]hemicholinium-3 ([3H]HCh-3) and high-affinity [3H]choline uptake. Potassium depolarization caused a significant 2-fold increase in the specific binding of [3H]HCh-3 in slices of rat striatum in vitro. CaM antagonists, including trifluoperazine (TFP), W-5, W-7, promethazine and haloperidol, dose-dependently inhibited potassium depolarization-stimulated [3H]HCh-3 binding with IC50s of 20, 40, 70, 30 and 48 (microM), respectively. Scatchard analysis revealed that the inhibitory effect of TFP resulted from a decrease in Bmax but no change in Kd of [3H]HCh-3 binding. Potassium depolarization of slices also stimulated high-affinity [3H]choline uptake, which was completely inhibited by 10 microM TFP. These results are discussed in relation to the regulatory mechanisms of SDHACU.     

Audiological evaluation of the middle ear implant--temporal auditory acuity

Temporal auditory acuities provided by the middle ear implant (MEI) and by the hearing aid (HA) were compared in ten patients implanted with the MEI. Test sounds used in the experiment were tone bursts [rise and fall time: 25 msec, duration (t): 982-1000 msec, interval (T): 1000 msec] of 500 Hz or 2000 Hz at 70 dBSPL. A speaker was placed in front of a subject one meter apart. At first, the subject adjusted the gain control of outer unit at the most comfortable loudness level. He heard totally fifty times of the test sounds, which varied in duration each 2 msec difference between 982 and 1000 msec and were provided in random manner. Therefore, sound of each duration was given 5 times. The notch [= (T-t) x 2] of the test sounds recognized by the patients was regarded as time gap. Fifty percent discrimination thresholds, which were indicated by the gap times of the test sounds half identified, were obtained as the index of the temporal auditory acuity. Following the above test, performance of the HA was investigated by use of the same procedure. The results indicated that the temporal auditory acuities of the MEI were superior to those of the HA at 500 Hz (P less than 0.01) and 2000 Hz (P less than 0.001). When the intensity of the test sound at 2000 Hz decreased from 70 dBSPL to 60 or 50 dBSPL, the temporal auditory acuities of the MEI and the HA were almost depreciated together with consistent difference in the test of 6 patients (P less than 0.01, 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Adenovirus-mediated overexpression of a gene prevents hearing loss and progressive inner hair cell loss after transient cochlear ischemia in gerbils

The use of adenoviral vectors has recently provided a novel strategy for direct gene transfer into the cochlea. In this study, we assessed the utility of an adenoviral vector expressing glial-cell-derived neurotrophic factor (GDNF) in ischemia-reperfusion injury of the gerbil cochlea. The vector was injected through the round window 4 days before ischemic insult. The distribution of a reporter transgene was confirmed throughout the cochlea from the basal to the apical turn and Western blot analysis indicated significant upregulation of GDNF protein 11 days following virus inoculation. Hearing ability was assessed by sequentially recording compound action potentials (CAP), and the degree of hair cell loss in the organ of Corti was evaluated in specimens stained with rhodamine-phalloidin and Hoechst 33342. On the seventh day of ischemia, the CAP threshold shift and inner hair cell loss were remarkably suppressed in the Ad-GDNF group compared with the control group. These results suggest that adenovirus-mediated overexpression of GDNF is useful for protection against hair cell damage, which otherwise eventually occurs after transient ischemia of the cochlea.     

Change of platelet activation markers using flow cytometry in patients with hematology/oncology disorders after transfusion

In spite of the frequent need of platelet transfusions, there is limited information on the association of platelet activation markers, in transfused patients with hematology/oncology disorders, with platelet function using flow cytometry. The goal of this study was to evaluate the changes of PAC-1 binding and CD62P expression, with or without agonists in patients after transfusions. Twenty-eight whole blood samples were obtained from 24 patients admitted to the department of Hematology & Oncology and transfused with platelets; these samples were compared to 30 healthy controls. Whole blood samples, either with or without agonists, such as 20 microM adenosine diphosphate (ADP) or 100 microM thrombin receptor activating peptide (TRAP), were stained with the fluorescein conjugated monoclonal antibodies PAC-1 or CD62P. Then, the percent expression for each marker was analysed using flow cytometry. ADP and TRAP induced an increased percentage of CD62P expression and PAC-1 binding after platelet transfusions compared to the samples studied before transfusion, and these findings were lower than those of the healthy controls. However, the expression of platelets without the agonists was not significantly changed, despite the transfusions. Therefore, agonist-induced platelet activation markers, studied by flow cytometry, appear to be more useful for the evaluation of platelet function after transfusions than platelet activation markers without agonists.     

Novel complex class 1 integron bearing an ISCR1 element in an Escherichia coli isolate carrying the blaCTX-M-14 gene

This work identifies an ISCR1-related bla(CTX-M-14) gene, which has never been reported before, from a clinical isolate of Escherichia coli. The bla(CTX-M-14) gene was preceded by an ISCR1 element that was followed by a class 1 integron containing three different insert gene cassettes, i.e., dfrA12, orfF, and aadA2.     

Overexpression of piccolo C2A domain induces depression-like behavior in mice

Piccolo is one of the components of the active zone at chemical synapses and regulates the transport of synaptic vesicles. The piccolo C2A domain is an important calcium sensor and binds with phosphatidylinositol or synaptotagmin-1. Recently, clinical studies suggested that a single nucleotide polymorphism in the piccolo C2A domain might be a causal risk factor for major depression. To clarify the association of piccolo with depression, we produced a transgenic mouse overexpressing the C2A domain of piccolo, and investigated the behavior of these mice. The mice exhibited depression-like behavior in both forced swim and tail suspension tests, suggesting that piccolo might regulate the depressive behavior.     

Docetaxel inhibits bone resorption through suppression of osteoclast formation and function in different manners

Osteoclasts are formed from the monocyte-macrophage lineage in response to receptor activator of nuclear factor κB ligand (RANKL) expressed by osteoblasts. Bone is the most common site of breast cancer metastasis, and osteoclasts play roles in the metastasis. The taxane-derived compounds paclitaxel and docetaxel are used for the treatment of malignant diseases, including breast cancer. Here we explored the effects of docetaxel on osteoclastic bone resorption in mouse culture systems. Osteoclasts were formed within 6 days in cocultures of osteoblasts and bone marrow cells treated with 1,25-dihydroxyvitamin D₃ plus prostaglandin E₂. Docetaxel at 10⁻⁸ M inhibited osteoclast formation in the coculture when added for the entire culture period or for the first 3 days. Docetaxel, even at 10⁻⁶ M added for the final 3 days, failed to inhibit osteoclast formation. Osteoprotegerin, a decoy receptor of RANKL, completely inhibited osteoclast formation when added for the final 3 days. Docetaxel at 10⁻⁸ M inhibited the proliferation of osteoblasts and bone marrow cells. RANKL mRNA expression induced by 1,25-dihydroxyvitamin D₃ plus prostaglandin E₂ in osteoblasts was not affected by docetaxel even at 10⁻⁶ M. Docetaxel at 10⁻⁶ M, but not at 10⁻⁸ M, inhibited pit-forming activity of osteoclasts cultured on dentine. Actin ring formation and l-glutamate secretion by osteoclasts were also inhibited by docetaxel at 10⁻⁶ M. Thus, docetaxel inhibits bone resorption in two different manners: inhibition of osteoclast formation at 10⁻⁸ M and of osteoclast function at 10⁻⁶ M. These results suggest that taxanes have beneficial effects in the treatment of bone metastatic cancers. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

GABAB receptor agonist baclofen improves methamphetamine-induced cognitive deficit in mice

In this study, we investigated the effects of GABA_A and GABA_B receptor agonists on the methamphetamine-induced impairment of recognition memory in mice. Repeated treatment with methamphetamine at a dose of 1 mg/kg for 7 days induced an impairment of recognition memory. Baclofen, a GABA_B receptor agonist, ameliorated the repeated methamphetamine-induced cognitive impairment, although gaboxadol, a GABA_A receptor agonist, had no significant effect. GABA_B receptors may constitute a putative new target in treating cognitive deficits in patients suffering from schizophrenia, as well as methamphetamine psychosis.